Pulschen Andre A, Bendia Amanda G, Fricker Ashwana D, Pellizari Vivian H, Galante Douglas, Rodrigues Fabio
Instituto de Química, Universidade de São PauloButantã, Brazil.
Departamento de Oceanografia Biológica, Instituto Oceanográfico, Universidade de São PauloButantã, Brazil.
Front Microbiol. 2017 Jul 14;8:1346. doi: 10.3389/fmicb.2017.01346. eCollection 2017.
Uncultured microorganisms comprise most of the microbial diversity existing on our planet. Despite advances in environmental sequencing and single-cell genomics, in-depth studies about bacterial metabolism and screening of novel bioproducts can only be assessed by culturing microbes in the laboratory. Here we report uncultured, or recalcitrant, microorganisms from an Antarctic soil sample, using relatively simple methods: oligotrophic media, extended incubation periods, observation under stereo microscopy, and selection of slow-growing bacteria. We managed to isolate several rare microorganisms belonging to infrequently isolated or recently described genera, for example , and Additionally, we obtained isolates presenting 16S rRNA sequence similarity ranging from 92.08 to 94.46% with any other known cultured species, including two distinct isolates from the class , that although common in Antarctic soils (as identified by metagenomics), was never reported to be isolated from such samples. Our data indicates that simple methods are still useful for cultivating recalcitrant microorganisms, even when dealing with samples from extreme environments.
未培养的微生物构成了地球上现存微生物多样性的大部分。尽管环境测序和单细胞基因组学取得了进展,但关于细菌代谢的深入研究和新型生物产品的筛选只能通过在实验室中培养微生物来评估。在这里,我们报告了使用相对简单的方法从南极土壤样本中分离出未培养的或难培养的微生物:贫营养培养基、延长培养时间、立体显微镜观察以及选择生长缓慢的细菌。我们成功分离出了几种属于罕见分离或最近描述的属的稀有微生物,例如 ,此外,我们还获得了与任何其他已知培养物种的16S rRNA序列相似性在92.08%至94.46%之间的分离株,包括来自 类的两个不同分离株,该类在南极土壤中很常见(通过宏基因组学鉴定),但从未报道过从此类样本中分离出来。我们的数据表明,即使处理来自极端环境的样本,简单的方法对于培养难培养的微生物仍然有用。